CA2930489A1 - Compositions based on methyl cyclodextrins for the treatment and/or prevention of diseases by increasing the hdl cholesterol level - Google Patents

Abstract

The invention relates to a novel use of a pharmaceutical composition comprising at least one methyl cyclodextrin having a molar substitution (MS) degree of between 0.05 and 1.5 in the treatment and/or prevention of diseases that can be treated and/or prevented by an increase in the HDL cholesterol level. The invention also relates to the use thereof in the treatment and/or prevention of atherosclerosis or complications relating to an atheroma, and/or diseases of the central nervous system. The pharmaceutical compositions of the invention can also be used to stimulate the reduction of circulating fatty acid and triglyceride levels and to reduce or prevent atheromatous plaques.

Description

COMPOSITIONS BASED ON METHYL-CYCLODEXTRINS FOR
TREATMENT AND / OR PREVENTION OF DISEASES BY INCREASE
CHOLESTEROL-HDL RATE.
The present invention relates to pharmaceutical compositions allowing to increase the HDL cholesterol level in an individual. The present invention concerned more specifically the use of a pharmaceutical composition in the treatment and / or the prevention of diseases that can be treated and / or prevented by one increase in HDL cholesterol, or in the treatment and / or prevention of atherosclerosis or complications related to atheroma, or system central nervous system.
BACKGROUND OF THE INVENTION
Cholesterol is a lipid of the family of sterols playing a central role in many biochemical processes.
It is a major component of animal cell membranes that contributes to their stability and the maintenance of their structure by interposing phospholipids. Indeed, cholesterol stiffens the membrane by preventing its gelation because it avoids the crystallization of fatty acids and also decreases the permeability Membrane water soluble molecules. By intercalating in membranes, cholesterol allows also the formation of lipid rafts which are essential areas to anchoring functional proteins.
In addition, cholesterol is also found in neurons where it allows the synthesis of neurotransmitters by exocytosis and therefore the spread of the nerve message. he is directly involved in the formation of beta-amyloid peptides, and therefore in the pathogenesis of Alzheimer's disease. An association between atherosclerosis and the Alzheimer's disease has also been demonstrated (Mikael Simons et al., Cholesterol depletion inhibitors the generation of b-amyloid in hippocampal neurons, Proc. Natl.
Acad. Sci. USA;
Flight. 95, pp. 6460-6464, May 1998, Neurobiology.) WO 2015/087016

2 PCT / FR2014 / 053309 The metabolism of cholesterol is also a precursor of many molecules as for example steroid hormones (cortisol, cortisone and aldosterone), hormones sex steroids (progesterone, estrogen and testosterone), vitamin D3, the heme A, prenylated or farnesylated proteins, ubiquinone or coenzyme Q10, dolichol, the NF kappa B nuclear factor, Tau protein and bile salts.
Cholesterol is carried in the blood by very roles different, namely LDL lipoproteins (low density lipoproteins) and the HDL lipoproteins (high density lipoproteins).
HDL-cholesterol is considered protective against diseases cardiovascular and is frequently referred to as good cholesterol.
LDL carrying cholesterol enter the arterial wall and then are picked up by monocytes and macrophages residing in this arterial wall as well as by the cells smooth muscles. This results in an excessive lipid load of macrophages and cells smooth muscles that then turn into foam cells at the origin process atherosclerosis and corresponding to the first stage of the formation of plate atheroma. It is about bad cholesterol.
Atheroma is defined by the World Health Organization as a "association of reshaping the intima of the large and medium-sized arteries consisting of Focal accumulation of lipids, complex carbohydrates, blood and deposits limestone, with reworking of the media. "Atheroma is at the origin of the majority of diseases cardiovascular disease and is the leading cause of morbidity and mortality among country industrialized. Atheroma begins with the formation of a lipidic streak, simple deposit of fat, linear and inconsequential to the flow, located between endothelium and media of the artery. With time, this streak can grow, become loaded with lipids, fibrogenic, in platelets and other blood cells and calcium to form the plate of atheroma. It becomes more or less important and can decrease enough the caliber of the artery to decrease its flow.
There are two types of complications: the first, of long evolution, is due to his slow growth, annoying more and more the passage of blood to prevent it totally WO 2015/087016

3 PCT / FR2014 / 053309 by obstruction. The second, fast and responsible for acute complications, consists of the lesion or rupture of the endothelium: the breach formed is then obstructed by one aggregation of platelets and the formation of a blood clot that can quickly completely obstruct the ship. This clot can also be detach and obstruct further downstream. The plate can also come off partially and obstruct also the artery, or more rarely, release its contents and make an embolism of cholesterol. Finally, the dilation of the arterial wall induced by volume increase plaque may result in the formation of an aneurysm, with the risk of break.
The most used pharmaceutical class for preventing atheroma are the statins, which aim to reduce LDL-cholesterol and circulating lipids, by complement of a adapted diet. However the interest and utility of statins are since some times strongly criticized by some researchers and clinicians who claim that statins, after analysis of controversial statistical tests because contradictory, do not bring not to patients the expected therapeutic benefit. To this challenge are added the Side effects harmful, and even sometimes dramatic, that have been identified and have led in some case of withdrawal from the market.
The other medications proposed today are:
platelet antiaggregants such as aspirin or clopidogrel, which are supposed to decrease the formation of clots from the atheroma plaque;
- antihypertensive drugs such as inhibitors of the enzyme conversion of angiotensin.
The existing pharmaceutical treatments are therefore aimed at only to to reduce the risks related to atheroma. None of the existing drugs on the market does directly attacks the atheroma plaque.
When life-threatening is at stake, the treatment is surgical or Endovascular. It aims to restore the arterial lumen, to irrigate the territory deprived of oxygen or to delete the arterial aneurysm. These techniques include angioplasty, clearing by endarterectomy, bypass.

WO 2015/087016

4 PCT / FR2014 / 053309 Finally, the main way to fight against atheroma and its complications keep staying behavioral today: cessation of tobacco, development of physical activity, control of blood pressure, correction of dyslipidemia, balance of diabetes, diet.
Thus, it is clear that there is clearly and for a long time a need not satisfied with treatment and / or prevention of atheroma.
However, various approaches have been tried without any new solution could not be detention. HDL cholesterol in particular began to be studied in 1975 when researchers have highlighted the relationship between HDL-cholesterol levels high and the decreased incidence of cardiovascular disease (Rye KA 2013. High density lipoprotein structure, function, and metabolism: a new Thematic Series. J.
Lipid Res.
54: (8) 2031-2033). The positive results of these studies encouraged the development of treatments aimed at increasing HDL-cholesterol levels, among which we will remember inhibitors of the CETP enzyme (protein transfer protein esters cholesterol) that have been the first agents developed and evaluated on a large scale, specifically in this goal. However, none of these treatments reduced the occurrence of accidents Cardiovascular.
Finally, there is still no drug on the market to induce effectively increase the HDL-cholesterol level in the patient, especially because the mechanisms involved are still far from being elucidated. In In particular, there is no drug on the market capable of effectively and actively against diseases that can be treated and / or prevented by one increase in HDL-cholesterol In addition, there is a need constant of drug capable of combating atherosclerosis, particularly against formation of the atheroma plaque.
SUMMARY OF THE INVENTION
It is the merit of the Claimant to have discovered that compositions methyl-cyclodextrin-based pharmaceutical products with a degree of substitution molar range of between 0.05 and 1.5 allows to induce a increase in WO 2015/087016

5 PCT / FR2014 / 053309 HDL-cholesterol levels, especially HDL-cholesterol plasma, and are therefore extremely interesting for the treatment or prevention of diseases likely to be treated and / or prevented by an increase in the rate of cholesterol-HDL, including atherosclerosis or complications related to atheroma, or some diseases of the central nervous system.
The object of the present invention is therefore to propose a composition pharmaceutical comprising at least one methyl-cyclodextrin having a degree of molar substitution between 0.05 and 1.5 for its use in the treatment and / or prevention of diseases that can be treated and / or prevented by an increase in rate HDL cholesterol.
Another object of the present invention is to provide a composition pharmaceutical comprising at least one methyl-cyclodextrin having a degree of molar substitution between 0.05 and 1.5 for its use in the treatment and / or prevention of atherosclerosis or complications related to atheroma, or system central nervous system.
Preferably, the compositions of the invention are used in the treatment and / or prevention of atherosclerosis or complications related to atheroma. The complications related to atheroma typically include ischemia, for example ischemia of myocardium, coronary heart disease, angina pectoris, acute coronary myocardial infarction, mesenteric infarction, stroke cerebral, aneurysm, or arterial disease of the lower limbs and their consequences (Consequences related to hypopoxia / ischemia, for example diabetes consecutive to atherosclerosis).
In another preferred embodiment, the compositions of the invention used in the treatment and / or the prevention of diseases of the central nervous system are chosen among the disease Alzheimer's disease, Parkinson's disease and lysosomal diseases affecting the system central nervous system. Preferentially, the lysosomal diseases affecting the the nervous system are selected from Niemann-Pick disease, preferentially the disease Niemann-Pick type A, Niemann-Pick type B disease, or Niemann-Pick type C.

WO 2015/087016

6 PCT / FR2014 / 053309 Methyl-cyclodextrin used in the pharmaceutical compositions of the invention preferably has a degree of molar substitution of between 0.2 and 1,2, and more preferably between 0.4 and 0.9. Advantageously, the degree of molar substitution is between 0.6 and 0.8. In one embodiment particular, the methyl-cyclodextrin is a methyl-13-cyclodextrin. In another mode of production In particular, the methyl-cyclodextrin is substituted on the hydroxyl borne by C2 carbon glucopyranose units, or by the C3 and / or C6 carbons of the units glucopyranoses or by a combination of carbons C2, C3 and / or C6, preferably C2 and C6 units glucopyranose.
Preferably, the methyl-cyclodextrin compositions comprise one or many methyl-13-cyclodextrins selected from the group consisting of cyclodextrins substituted on the hydroxyl carried by the carbon C2 of the units glucopyranoses, hydroxyl substituted methylcyclodextrins by the C3 and / or C6 carbons of glucopyranose units, methyl-13-cyclodextrins substituted on the hydroxyl borne by the carbons C2, C3 and / or C6, preferably C2 and C6 units glucopyranoses and said methyl-13-cyclodextrins having a degree of substitution molar range from 0.6 to 0.8.
Advantageously, the methyl-cyclodextrin compositions comprise at least minus 50, 60, or 75% of methyl substituted on the hydroxyl carried by the carbon C2 units glucopyranose.
Optionally, the compositions comprising at least one methyl-cyclodextrin also include a cyclodextrin, especially p. ¨cyclodextrin, no substituted and / or a cyclodextrin, in particular p. ¨cyclodextrin, substituted by groups sulfobutyl ether (SBE-) and hydroxypropyl (HP-), preferably with a degree of molar substitution between 0.05 and 1.5.
The pharmaceutical compositions used in the context of this invention can further comprising at least one additional active agent.
The additional active agent is preferably an active agent used in the treatment atherosclerosis or complications related to atheroma, preferably, WO 2015/087016

7 PCT / FR2014 / 053309 - a statin, a platelet antiaggregant or anticoagulant agent, preferably selected aspirin, clopidogrel, new oral anticoagulants such as dabigatran, apixaban, rivaroxaban, and anti-vitamin K
an antihypertensive agent, preferably selected from inhibitors of angiotensin converting enzyme such as perindopril, captopril, enalapril, the lisinopril or ramipril, angiotensin II receptor antagonists as the losartan, valsartan, or candesartan, and / or among beta-blockers as acebutolol, labetalol, nadolol, oxprenolol, penbutolol, pindolol, or propranolol - or a combination of these.
Preferably, the additional active agent is an antagonist of the receiver to angiotensin II, such as losartan, valsartan, or candesartan.
The pharmaceutical compositions according to the invention are likely to be administered orally, parenterally, or cutaneously or mucosally.
In a particular embodiment, the compositions of the invention are in addition used to promote lowering of triglyceride and acid levels circulating fat.
In another particular embodiment, the compositions of the invention are used to reduce or prevent atheromatous plaque.
In yet another particular embodiment, the compositions used in the invention are free or substantially free of vesicles from phospholipids.
A further object of the invention is the non-use therapeutic of a composition comprising at least one methyl-cyclodextrin having a degree of molar substitution between 0.05 and 1.50, preferably between 0.60 and 0.80, to lower the triglyceride level and / or to reduce the concentration of fatty acids free and / or to increase the HDL-cholesterol level.

WO 2015/087016

8 PCT / FR2014 / 053309 DETAILED DESCRIPTION OF THE INVENTION
The inventors have identified a new use of a composition pharmaceutical comprising at least one methyl-cyclodextrin having a degree of molar substitution between 0.05 and 1.5 in the treatment and / or prevention of diseases likely to be treated and / or prevented by an increase in cholesterol HDL and / or a reduction or prevention of atheromatous plaques.
In particular, the inventors have surprisingly discovered that the composition of the effect of the invention was to increase the HDL cholesterol level in a individual, and In addition, lowering the level of triglycerides and circulating fatty acids and reduce Atheroma plaques. The inventors have thus demonstrated that the composition of the invention thanks to its different properties can be used especially in the treatment and / or prevention of diseases related to overloading, and / or storage, and / or the accumulation of cholesterol in the tissues, as well as their consequences. In particular, these diseases include atherosclerosis or atheroma-related complications, and diseases of central nervous system, including Alzheimer's, Parkinson's or Parkinson's disease Niemann Pick type C.
The invention therefore relates to a pharmaceutical composition comprising at least a methyl-cyclodextrin having a degree of molar substitution between 0.05 and 1.5 for use in the treatment and / or prevention of diseases likely to be treated and / or prevented by an increase in HDL-cholesterol particular atherosclerosis or complications related to atheroma, and / or diseases of the system central nervous system, including Alzheimer's, Parkinson's or Niemann's Pick type C. It also relates to a pharmaceutical composition comprising at least less a methyl-cyclodextrin having a degree of molar substitution understood between 0.05 and 1.5 for its use in the treatment and / or prevention of diseases likely to be treated and / or prevented by the reduction of atheromatous plaques or by prevention the formation of atheroma plaques.
The invention also relates to a pharmaceutical composition comprising at least a methyl-cyclodextrin having a degree of molar substitution between 0.05 and 1.5 for its use in the treatment and / or prevention of atherosclerosis or WO 2015/087016

9 PCT / FR2014 / 053309 complications related to atheroma, and / or diseases of the nervous system central, especially Alzheimer 's, Parkinson' s or Niemann Pick type C.
The invention also relates to methods or methods employing a pharmaceutical composition comprising at least one methyl-cyclodextrin such than defined in the present invention for its administration in a quantity therapeutic effective in an individual suffering from an illness that can be treated and / or prevented by an increase in HDL cholesterol and / or by a reduction or prevention Atheroma plaques, particularly atherosclerosis or complications linked to a atheroma, and / or diseases of the central nervous system, including Alzheimer's, Parkinson's or Niemann Pick type C.
The invention also relates to the use of a pharmaceutical composition comprising at least one methyl-cyclodextrin as defined in the this application for the preparation of a medicament for treating and / or preventing diseases likely to be treated and / or prevented by an increase in cholesterol HDL and / or a reduction or prevention of atheromatous plaques, preferably atherosclerosis or complications related to atheroma, and / or diseases of the nervous system central, especially Alzheimer 's, Parkinson' s or Niemann Pick type C.
Methyl cyclodextrins Cyclodextrins are cyclic oligosaccharides from the degradation enzymatic starch. The three most natural cyclodextrins current ones consist of 6, 7 or 8 units aD-glucopyranose in chair configuration connected between they by links a-1,4. They are more commonly called a, p, or cyclodextrin respectively. Their three-dimensional structure appears in the form of a cone truncated outside which are the hydroxyl groups representing the party highly hydrophilic cyclodextrins. The interior of the cone or the cavity of cyclodextrins is constituted by the hydrogen atoms carried by the C3 and C5 carbons as well as by the oxygen atoms participating in the glycosidic bond, their giving thus an apolar character. Cyclodextrins having a part hydrophilic exterior and a hydrophobic cavity are generally used for their ability to encapsulate hydrophobic compounds and, therefore, for their role of protector and solubilizer of WO 2015/087016

10 PCT / FR2014 / 053309 hydrophobic active substances. They are thus classically found in areas of agribusiness, but also in galenics where they are used as excipient in pharmaceutical formulations administered orally or in formulations cosmetics administered topically.
In order to improve the aqueous solubility of natural cyclodextrins, numerous derivatives have been synthesized by grafting different groups on the functions hydroxyls. The glucopyranose units of cyclodextrins do indeed comprise each 3 reactive hydroxyl groups, which are carried by the carbons C2, C3 and C6.
Examples of derivatives that may be mentioned are hydroxypropyl cyclodextrins, methyl cyclodextrins and sulfated derivatives of cyclodextrin. Fromming and Szetli have particularly shown in the case of methyl-cyclodextrins that the increase in degree of methylation promoted solubilization to a degree of substitution molar equal to 2, and that beyond the solubilization decreased.
Some authors have also been interested in cyclodextrins and their derivatives otherwise than by their role as a pharmaceutical excipient.
For example, patent application WO 02/43742 describes the role of cyclodextrins or hydroxypropyl-cyclodextrins in the metabolism of cholesterol, and especially for promote the efflux of cholesterol macrophages. In this application, authors recall that there are two mechanisms of actions of cyclodextrins. The cyclodextrins high affinity for cholesterol act directly by complexing the cholesterol membrane of macrophages. In contrast, low cyclodextrins affinity for the cholesterol act as a catalyst for the efflux of cholesterol, cellular membrane to an extracellular acceptor which is a phospholipid vesicle. In this last case, the efflux of cholesterol can be done only in the presence of another element that is an exogenous acceptor phospholipid vesicle. So, if effects pharmacological on cholesterol are mentioned in the prior art, the therapeutic interest of the cyclodextrins remains highly questionable.
Indeed, high affinity cyclodextrins, by their activity in the complexation of cholesterol levels, would have needed to be administered in considerable, WO 2015/087016

11 PCT / FR2014 / 053309 would not have been pharmaceutically acceptable and would have resulted in problems toxicity because of the large amount of solubilized cholesterol.
As for low-affinity cyclodextrins, the need for co-administer with phospholipid vesicles makes it a far too complex technology in view of of the current knowledge, particularly because phospholipid vesicles are very unstable.
The use of methyl-cyclodextrins among other active agents has been suggested in the treatment of atherosclerosis (WO 2006/032905). In particular, this request for patent suggests that methyl-cyclodextrins are able to inhibit between LDL
oxidized and monocytes. The use of methyl-13-cyclodextrins has been suggested in methods to modulate the inflammatory response by altering the levels plasma cholesterol (US 2008/0032925). Methyl-13-cyclodextrin used in this application is a methyl-13-cyclodextrin marketed by Sigma Aldrich and which presents a degree molar substitution well above 1.5.
In the present invention, the inventors use cyclodextrins particulars acting as an active ingredient for the purpose of increasing significantly the plasma HDL cholesterol levels while minimizing the related disadvantages to toxicity in a patient. The compositions of the invention allow to exclude problems of toxicity due to their particular characteristic, ie, a degree of substitution, in particular of methylation, relatively low affinity for cholesterol.
Surprisingly, and despite their low affinity for cholesterol, methyl-cyclodextrins, and in particular methyl-13-cyclodextrins, used in accordance with the present invention, do not require the addition of vesicles of phospholipids for exert their effect.
The results obtained by the inventors suggest a different mode of action for these specific methyl-cyclodextrins, which would not go through complexation cholesterol, nor by catalysis of transport to phospholipid vesicles.

WO 2015/087016

12 PCT / FR2014 / 053309 In a very original way, these methyl-cyclodextrins seem to promote the efflux of cholesterol in the form of HDL-cholesterol, and not in a form complexed with cyclodextrins, or phospholipid vesicles.
This is why the compositions according to the present invention are free or substantially free of phospholipid vesicles. Subtantiellement free here means that the compositions comprise less than 10% of phospholipids in weight relative to the composition, preferably less than 5% by weight and way particularly preferred, less than 2, 1 or 0.5% by weight. Subtantiellement free can also be defined by a ratio in molar concentration between the number of cyclodextrin and that of phospholipids which is greater than 100, 1000, or 10 000.
The present invention therefore relates to a new use of a composition pharmaceutical composition comprising at least one methyl-cyclodextrin having a degree of molar substitution between 0.05 and 1.5 in the treatment and / or prevention of diseases that can be treated and / or prevented by an increase in rate HDL cholesterol and / or by reduction or prevention of atheromatous plaque.
The invention also relates to a new use of a composition pharmaceutical comprising at least one methyl-cyclodextrin having a degree of molar substitution between 0.05 and 1.5 in the treatment and / or prevention of atherosclerosis or complications related to atheroma, or diseases of the central nervous system, in especially Alzheimer's, Parkinson's or Niemann Pick type C. means by molar degree of substitution (MS) the number of substituted hydroxyls, especially by a methyl group, per glucopyranose unit. Note that the degree of substitution Molar (MS) is different from the degree of molecular substitution (DS) that corresponds to number of hydroxyls substituted, in particular by a methyl group, by molecule of cyclodextrin and therefore taking into account the number of glucopyranose units constituting the methyl-cyclodextrin.
MS can be determined in the present invention by Magnetic Resonance Nuclear proton (NMR), or by mass spectrometry (spectrometry of mass by electrospray ionization (ESI-MS) or mass spectrometry matrix assisted laser desorption / ionization (MALDI-MS)). Although these techniques WO 2015/087016

13 PCT / FR2014 / 053309 are well known to those skilled in the art, optimal conditions of determination of MS methyl-cyclodextrins according to the invention are particularly well described in the reference thesis of JACQUET Romain. Hydrophilic cyclodextrins:
characterization and study of their enantioselective and complexing properties. Using the liquid chromatography and mass spectrometry. Thesis Chemistry and physicochemistry of compounds of biological interest. University of Orleans, 2006.
especially available on: http: // tel. open archive .fr / docs / 00/18/55/42 / PDF / jacquet.pdf (accessed 27.11.2013). , in particular Chapter 2, Part B (pages 59 to 83).
Preferably, the MS is determined by NMR, according to the following method:
measures are 25 C on a 250 MHz DPX-type device Advance (Bruker, Rheinstetten, Germany). Calibration is performed with signal D20. The samples of methyl cyclodextrin according to the invention, and native cyclodextrin, that is to say no methylated, are prepared at a concentration of 5 mg in 0.75 mL of D20. The solutions are evaporated to dryness under a stream of nitrogen and then reconstituted in 0.75 mL of D20. This operation is repeated twice to ensure a total exchange of protons functions hydroxyl. The MS is calculated from the difference in integration between the spectrum of cyclodextrin and that of methyl-cyclodextrin according to the invention. A ghost typical is shown in Figure 1.
It should be noted that the methyl cyclodextrin used in accordance with the invention, although which may correspond to a pure product, generally corresponds to a mixture of methyl cyclodextrins of different structures. This is the case, for example, with the product marketed by the Applicant under the name KLEPTOSE CRYSMEB, which presents in particular the physicochemical properties as determined in the thesis JACQUET Romain, in particular in Chapter 2, Part B (pages 59 to 83).
As a result, the measured MS is in this case an average of the substitutions that are happening of all the glucopyranose units in the whole mixture of methyl-cyclodextrins.
This mixture may in particular contain residual native cyclodextrin, that is to say, no methylated, but which are usually in negligible quantities, particular less than 1% by dry weight relative to the total dry weight of methyl-cyclodextrin preferably less than 0.5%, preferentially still less than 0.1%.

WO 2015/087016

14 PCT / FR2014 / 053309 In the context of the invention, the compositions comprise at least one methyl cyclodextrin having a degree of molar substitution of between 0.05 and 1.5.
Advantageously, the methyl-cyclodextrin has an MS of between 0.1 and 1.4, preferably between 0.1 and 1.3, preferably between 0.2 and 1.2, preferably between 0.3 and 1.1, preferably between 0.3 and 1, preferentially between 0.5 and 0.9, preferably between 0.6 and 0.8, for example 0.7, especially 0.67. By example, the methyl-cyclodextrin can have an MS between 0.10 and 1.40, between 0.10 and 1.30, between 0.10 and 1.20, between 0.15 and 1.40, between 0.15 and 1.30, between 0.15 and 1.20, between 0.20 and 1.40, between 0.20 and 1.30, between 0.20 and 1.20, between 0.20 and 1.10, between 0.25 and 1.40, between 0.25 and 1.30, between 0.25 and 1.20, between 0.25 and 1.10, between 0.15 and 0.90, between 0.15 and and 0.80, between 0.25 and 1.00, between 0.25 and 0.90, between 0.25 and 0.80, between 0.30 and 1.40, 0.30 and 1.30, between 0.30 and 1.20, between 0.30 and 1.00, between 0.50 and 0.90, between 0.60 and 0.80.
Preferably, at least 50% of the methyl groups of methyl-cyclodextrin used in the context of the present invention are located at the level of the hydroxyl carried by the carbon C2 of the glucopyranose unit, preferably between 60 and 80%
typically of the order of 75%.
At the same time, other methyl groups are generally mostly located at the level of the hydroxyl borne by the carbon C3 and / or C6 of the unit glucopyranose.
The person skilled in the art knows how to determine the distribution of the groups methylated on the hydroxyls of the glucopyranose unit of methyl-cyclodextrin, for example by NMR.
Advantageously, the methyl-cyclodextrin used in the context of this invention has 7 α-glucopyranose units. It is therefore a méthylecyclodextrine.
In a particular embodiment, methyl-cyclodextrin is a méthy1-13-cyclodextrin and has an MS of between 0.05 and 1.5, preferably including between 0.1 and 1.4, preferably between 0.1 and 1.3, preferably between 0.2 and 1.2, preferably between 0.3 and 1.1, preferably between 0.4 and 1, preferably between 0.5 and 0.9, preferably between 0.6 and 0.8, for example 0.7, in particular 0.67. By for example, the methyl-cyclodextrin may have an MS of between 0.10 and 1.40, between 0.10 WO 2015/087016

15 PCT / FR2014 / 053309 and 1.30, between 0.10 and 1.20, between 0.15 and 1.40, between 0.15 and 1.30, between 0,15 and 1,20, between 0.20 and 1.40, between 0.20 and 1.30, between 0.20 and 1.20, between 0.20 and 1.10, between 0.25 and 1.40, between 0.25 and 1.30, between 0.25 and 1.20, between 0.25 and 1.10, between 0.25 and 1.00, between 0.25 and 0.90, between 0.25 and 0.80, between 0.30 and 1.40, between 0.30 and 1.30, between 0.30 and and 1.20, between 0.30 and 1.00, between 0.50 and 0.90, between 0.60 and 0.80.
Methyl-cyclodextrin may be substituted on the hydroxyl borne by the carbon C2 of glucopyranose units, or by the C3 and / or C6 carbons of the units glucopyranoses, or by a combination of the C2, C3 and / or C6 carbons, preferably C2 and C6 units glucopyranose.
In another particular embodiment, methyl-cyclodextrin is a methyl cyclodextrin, preferably a methylcyclodextrin, at least 50% of which methyl groups are located at the level of the hydroxyl carried by the carbon C2 of the glucopyranose unit, preferably between 60 and 80%, typically from the order of 75%, and has an MS of between 0.05 and 1.5, preferably between 0.1 and 1.4 preferably between 0.1 and 1.3, preferably between 0.2 and 1.2, preferably between 0.3 and 1.1, preferably between 0.4 and 1, preferentially between 0.5 and 0.9, preferably between 0.6 and 0.8, for example 0.7, especially 0.67. By example, the methyl-cyclodextrin can have an MS between 0.10 and 1.40, between 0.10 and 1.30, between 0.10 and 1.20, between 0.15 and 1.40, between 0.15 and 1.30, between 0.15 and 1.20, between 0.20 and 1.40, between 0.20 and 1.30, between 0.20 and 1.20, between 0.20 and 1.10, between 0.25 and 1.40, between 0.25 and 1.30, between 0.25 and 1.20, between 0.25 and 1.10, between 0.25 and 1.00, between 0.25 and and 0.90, between 0.25 and 0.80, between 0.30 and 1.40, between 0.30 and 1.30, between 0.30 and 1.20, 0.30 and 1.00, between 0.50 and 0.90, between 0.60 and 0.80.
In a preferred embodiment, the methyl-cyclodextrin composition comprises one or more methyl-13-cyclodextrins selected from the group consisting of diced methylcyclodextrins substituted on the hydroxyl carried by the C2 carbon of the units glucopyranoses, hydroxyl substituted methylcyclodextrins speak C3 and / or C6 carbon of glucopyranose units, methylcyclodextrins substituted on the hydroxyl borne by the carbons C2, C3 and / or C6, preferably C2 and C6 units glucopyranoses and having an MS of between 0.05 and 1.5, preferentially between 0.1 and 1.4, preferably between 0.1 and 1.3, preferably between 0.2 and 1,2, WO 2015/087016

16 PCT / FR2014 / 053309 preferably between 0.3 and 1.1, preferably between 0.4 and 1, preferably between 0.5 and 0.9, preferably between 0.6 and 0.8, for example 0.7, in particular 0.67. By for example, the methyl-cyclodextrin may have an MS of between 0.10 and 1.40, between 0.10 and 1.30, between 0.10 and 1.20, between 0.15 and 1.40, between 0.15 and 1.30, between 0,15 and 1,20, between 0.20 and 1.40, between 0.20 and 1.30, between 0.20 and 1.20, between 0.20 and 1.10, between 0.25 and 1.40, between 0.25 and 1.30, between 0.25 and 1.20, between 0.25 and 1.10, between 0.25 and 1.00, between 0.25 and 0.90, between 0.25 and 0.80, between 0.30 and 1.40, between 0.30 and 1.30, between 0.30 and and 1.20, between 0.30 and 1.00, between 0.50 and 0.90, between 0.60 and 0.80. Preferably, the composition of methyl cyclodextrins comprises at least 50, 60, or 75% of methyl substituted on the hydroxyl borne by the carbon C2 of the glucopyranose units.
As mentioned above, the methyl-cyclodextrin according to the invention can be a mixed. Mass spectrometry analysis of KLEPTOSE CRYSMEB, who is a methyl-13-cyclodextrin, reveals in particular that it is a polydispersed product, consisting of seven major groups of methyl-cyclodextrins, which distinguish by their DS. This DS, which in theory can vary from 0 to 21 for a methyl-13-cyclodextrin, varies from 2-8 in the product KLEPTOSE CRYSMEB.
Advantageously, the compositions of the invention comprise a mixture of methyl cyclodextrins comprising at least 50, 60, 70, 80 or 90% of methyl cyclodextrins having an MS of 0.2 to 1.2. Preferably at least 40, 50, 60, 70, 80 or 90% of methyl-cyclodextrins have an MS of between 0.3 and 1.1. Of preferably minus 30, 40, 50, 60, 70, 80 or 90% methyl-cyclodextrins have an MS
understood between 0.5 and 0.9. Even more preferably, at least 25, 30, 40, 50, 60, 70, 80 or 90%
methyl-cyclodextrins have an MS of between 0.6 and 0.8.
The methyl-cyclodextrin compositions may be optionally prepared in adding different methyl-cyclodextrins with defined MS to get some compositions as defined in the present invention or they can to be obtained as a result of the synthesis of these.
Thus, in another particular embodiment, the composition of methyl-cyclodextrins, preferably methyl-13-cyclodextrins, has the profile of substitution, expressed in molar percentages, according to:

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17 PCT / FR2014 / 053309 0 to 5% of methyl-13-cyclodextrins comprise 2 methyl groups (DS of 2);
o 5 to 15% of methylcyclodextrins comprise 3 methyl groups (DS of 3);
20 to 25% of methyl-13-cyclodextrins comprise 4 groups methyls (DS of 4);
25 to 40% of methyl-13-cyclodextrins comprise 5 groups methyls (DS of 5);
15 to 25% of methyl-13-cyclodextrins comprise 6 groups methyls (DS of 6);
o 5 to 15% of methylcyclodextrins comprise 7 methyl groups (SD of 7);
0 to 5% of methyl-13-cyclodextrins comprise 8 methyl groups (DS of 8);
the total sum being generally of the order of 100%, although the composition can may contain traces of methyl-cyclodextrins of different DS, as well as traces of native cyclodextrin, that is to say unmethylated.
The substitution profile can be determined by any well-known technique of the man of the trade, for example by ESI-SM or MALDI-TOF-SM. The optimal conditions of determination of the substitution profile by these two methods are widely discussed in Romain JACQUET's thesis cited above, in Chapter 2, Part B, points 11.3 and 11.2 (pages 67 to 82) and Appendix II.
In a preferred embodiment, the methyl-cyclodextrin composition, of preferably methyl-13-cyclodextrins, is such that at least 50% of the groups are located at the level of the hydroxyl carried by the C2 carbon of the units glucopyranoses, preferably between 60 and 80%, typically of the order of 75%, and who presents the substitution profile, expressed in molar percentages, following :
0 to 5% of methyl-13-cyclodextrins comprise 2 methyl groups (DS of 2);
o 5 to 15% of methylcyclodextrins comprise 3 methyl groups (DS of 3);

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18 PCT / FR2014 / 053309 20 to 25% of methyl-13-cyclodextrins comprise 4 groups methyls (DS of 4);
25 to 40% of methyl-13-cyclodextrins comprise 5 groups methyls (DS of 5);
15 to 25% of methyl-13-cyclodextrins comprise 6 groups methyls (DS of 6);
o 5 to 15% of methylcyclodextrins comprise 7 methyl groups (SD of 7);
0 to 5% of methyl-13-cyclodextrins comprise 8 methyl groups (DS of 8);
the total sum being generally of the order of 100%, although the composition can may contain traces of methyl-cyclodextrins of different DS, as well as traces of native cyclodextrin, that is to say unmethylated.
It is also quite possible to consider proportions or isolate molecules or groups of molecules of methyl-cyclodextrins, especially in function of their DS.
Thus, in another particular embodiment, methyl-cyclodextrin is a methylcyclodextrin which has a DS selected from an integer ranging from from 2 to 8, in particular 2, 3, 4, 5, 6, 7 or 8.
In another preferred embodiment, methyl-cyclodextrin is a methyl cyclodextrin of which at least 50% of the methyl groups are located at the of the hydroxyl borne by the C2 carbon of the glucopyranose units, preferentially between 60 and 80%, typically of the order of 75%, and which has a DS selected from a number integer from 2 to 8, especially 2, 3, 4, 5, 6, 7 or 8.
In another particular embodiment, methyl-cyclodextrin, in particular the methylcyclodextrin, has an MS of between 0.1 and 0.3, in particular between 0.2 and 0.3, in particular between 0.20 and 0.30. In another particular embodiment, methyl-cyclodextrin, in particular methylcyclodextrin, has an MS between 0.3 and 0.5 in particular between 0.30 and 0.50. In another particular embodiment, methyl-cyclodextrin, in particular methylcyclodextrin, has an MS between 0.5 and 0.6 WO 2015/087016

19 PCT / FR2014 / 053309 especially between 0.50 and 0.60. In another particular embodiment, methyl-cyclodextrin, in particular methylcyclodextrin, has an MS between 0.6 and 0.7 in particular between 0.60 and 0.70. In another particular embodiment, methyl-cyclodextrin, in particular methylcyclodextrin, has an MS between 0.7 and 0.8 in particular between 0.70 and 0.80. In another particular embodiment, methyl-cyclodextrin, in particular methylcyclodextrin, has an MS between 0.8 and 0.9 especially between 0.80 and 0.90. In another particular embodiment, methyl-cyclodextrin, in particular methylcyclodextrin, has an MS between 0.9 and 1.1 especially between 0.90 and 1.10. In another particular embodiment, methyl-cyclodextrin, in particular methylcyclodextrin, has an MS between 1,1 and 1,2 especially between 1.10 and 1.20.
Generally, the methyl-cyclodextrin used in accordance with the invention presents a reducing sugar content of less than 1% by dry weight, preferentially less than 0.5%.
The composition of methylcyclodextrins according to the invention can be obtained speak process described in US Patent 6,602,860 Bi. An example of such composition is marketed by the group ROQUETTE FRERES under the commercial name KLEPTOSE CRYSMEB and has a molar degree of substitution of 0.7 or more precisely 0.67 methyl per unit of glucose.
Optionally, the composition according to the present invention may comprise in besides cyclodextrin, in particular cyclodextrin, unsubstituted and / or cyclodextrin, in particular is cyclodextrin, substituted by sulfobutyl ether groups (SBE-) hydroxyethyl, hydroxypropyl (HP-), carboxymethyl, carboxyethyl, acetyl, triacetyls, succinyls, ethyls, propyls, butyls, sulphates, preferably sulfobutyles and hydroxypropyls, preferably with a molar degree of substitution included between 0.05 and 1.5.
Optionally, the methyl-cyclodextrin according to the invention, in particular the méthy1-13¨

cyclodextrin, may be substituted by additional groups, in particular chosen from those listed before. It may, for example, be a methyl-I3¨cyclodextrine sulfated.

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20 PCT / FR2014 / 053309 In a separate embodiment, the present invention considers also the use for the treatment of diseases according to the present invention other derivatives of cyclodextrins, preferably cyclodextrin, having a degree of substitution molar according to the present invention, that is to say between 0.05 and 1.5 preferably between 0.1 and 1.4, preferably between 0.1 and 1.3, preferably between 0.2 and 1.2, preferably between 0.3 and 1.1, preferably between 0.4 and 1, preferably between 0.5 and 0.9, preferably between 0.6 and 0.8, by example 0.7. These cyclodextrin derivatives, preferably cyclodextrin, are substituted by a group selected from sulfobutyl ethers (SBE-) and hydroxypropyl (HP-). Preferably, these substitutions are mainly borne by the carbon C2 of glucopyranose units, typically at 50%, 60%, 70% or 80%.
In an alternative embodiment of the invention, methyl cyclodextrins such as defined in this application and composed of linked αD-glucopyranose units between they by A-1,4 bonds may be substituted in whole or in part by D-glucopyranoses linked together by α-1,6 bonds, in the compositions pharmaceutical compositions of the present invention.
Application In the context of the present invention, the pharmaceutical compositions comprising at least one methyl-cyclodextrin as defined in the present application are used in the treatment and / or prevention of to be treated and / or prevented by an increase in HDL cholesterol and / or reduction or prevention of atheromatous plaque, particularly atherosclerosis or complications related to atheroma, and / or diseases of the central nervous system, in particular the Alzheimer's disease, Parkinson's disease or Niemann Pick type C.
In the context of the present invention, the increase in the rate of HDL-cholesterol is measured in any type of biological fluid. It is preferentially the rate plasma HDL cholesterol. This HDL cholesterol level can be measured by all method known to those skilled in the art, for example by precipitation, or by direct methods in homogeneous phase using immunological techniques.
A

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21 PCT / FR2014 / 053309 assay of the protein portion (Apo Al protein) component HDL is also possible.
The HDL cholesterol level is generally expressed as the number of moles of cholesterol transported by high density lipoprotein (HDL) per liter of blood. The cholesterol-HDL is akin to good cholesterol as opposed to transported cholesterol by the LDL.
According to the invention, the diseases that can be treated and / or prevented by one increase in HDL cholesterol levels are all diseases whose symptoms and / or the causes disappear or are attenuated in a patient when the rate HDL-cholesterol is superior to HDL-cholesterol before treatment. Of preference, diseases are those that can be treated and / or prevented by increase HDL cholesterol level. By increase, can be heard a increase HDL cholesterol level of at least 5, 7.5, 10, 15 or 20%.
Diseases that can be treated and / or prevented by an increase the rate of HDL-cholesterol are preferentially the diseases associated with an overload, and / or storage, and / or accumulation of cholesterol in the tissues, as well as their consequences.
Examples of diseases that may be related to overload, and / or a storage and / or accumulation of cholesterol in the tissues, mention may be made of diseases Cardiovascular, vascular diseases, arterial diseases peripheral devices occlusives such as atherosclerosis or complications related to atheroma.
Other examples include diseases of the central nervous system likely to be related to overloading, and / or storage, and / or the accumulation of cholesterol in the tissues of the central nervous system. Such diseases include but not limited to Alzheimer's disease, Parkinson's disease, and diseases lysosomal affecting the central nervous system.

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22 PCT / FR2014 / 053309 Examples of lysosomal diseases include, but are not limited to disease Niemann-Pick, such as Niemann-Pick type A disease, Niemann-Pick type B, or Niemann-Pick type C.
A particular example is Niemann-Pick type C disease characterized by a non-esterified cholesterol accumulation in the Central Nervous System (CNS).
In a preferred embodiment, the disease is selected from sickness Alzheimer's disease, Parkinson's disease and Niemann-Pick type C disease.
Another particular example is Alzheimer's disease. accumulation of cholesterol in the brain can also be the cause of the disease Alzheimer's and an increase in HDL-cholesterol levels promotes its transport to the liver where he is degraded. Evidence suggests that dysfunction of Cholesterol metabolism in the brain and the vascular system is according to all probability closely involved in Alzheimer's disease.
The presence of too high a cholesterol level is often considered as a factor risk for the onset of cognitive impairment, dementia or Parkinson.
The involvement of cholesterol in increasing the risk of Parkinson has has also been mentioned in several studies.
In this context, a treatment whose effect is an increase in the rate of cholesterol-HDL that contributes to the transport of excess cholesterol to the liver where it is gradient takes all its meaning for this application. Indeed, a bundle of observations, data statistics and experiments at the molecular level lead to a role of more in addition to significant to cholesterol metabolism in the genesis of the sickness Alzheimer. In particular a convergence between atherosclerosis and sickness Alzheimer's disease (AD) and between Alzheimer's disease and Niemann-Pick type C is firmly established. This last rare disease is characterized by excessive accumulation of cholesterol in nerve cells.
Furthermore, the present invention also relates to diseases likely to be treated and / or prevented by reduction or prevention of the plates atheroma. By WO 2015/087016

23 PCT / FR2014 / 053309 reduction of atheroma plaques is preferably understood that the plates present a weaker surface in the presence of the treatment than in the absence of it.
In particular, the plate surface reduction of 10, 20, 30, 40 or 50%. Preferably, this reduction is at least 30 or 40%. The surface of atheromatous plaques can to be determined by any method known to those skilled in the art and in particular the methods medical imaging such as MRI (Magnetic Resonance Imaging).
By Atheromatous plaque prevention is understood to mean a slowdown in development or the formation of these plates.
Diseases that can be treated and / or prevented by a reduction or prevention atheromatous plaques, in particular cardiovascular diseases, diseases vascular diseases, and preferentially peripheral arterial diseases occlusives such than atherosclerosis or complications related to atheroma.
According to the invention, the diseases that can be treated and / or prevented are preferentially atherosclerosis or complications related to atheroma, disease Alzheimer's disease, Parkinson's disease or Niemann-Pick type C
advantageously, atherosclerosis or complications related to atheroma.
Complications related to an atheroma that are treated and / or prevented by use of a pharmaceutical composition comprising at least one methyl-cyclodextrin of the invention is in a nonlimiting manner ischemia, for example ischemia of myocardium, coronary heart disease, angina pectoris, acute coronary myocardial infarction, mesenteric infarction, stroke cerebral, aneurysm or arteriopathy of the lower limbs.
In another particular embodiment, the compositions of the invention used further comprise at least one additional active agent. The active agent additional is preferably selected from the group of active agents known by those skilled in the art to treat and / or prevent atherosclerosis or complications related to an atheroma.
The subjects to be treated are preferably humans or animals, preference of humans. They may have a proven illness (diagnosed or established) or to be at risk WO 2015/087016

24 PCT / FR2014 / 053309 to develop this disease. In particular, the present invention means by treatment a decrease in the causes or symptoms of a disease, the delay in the appearance of the disease, a slowdown in its development but also a healing.
Examples of active ingredients used in the treatment of atherosclerosis or complications related to atheroma include:
- statins;
antihypertensive agents, in particular:
o angiotensin converting enzyme inhibitors, for example the perindopril, captopril, enalapril, lisinopril, or ramipril;
o angiotensin II receptor antagonists, also known as last name sartans, such as losartan, valsartan, candesartan;
o beta-blockers, such as acebutolol, labetalol, nadolol, oxprenolol, penbutolol, pindolol, propranolol;
platelet antiaggregants or anticoagulants, in particular:
o asipirine;
o clopidogrel (PLAVIX10);
o new oral anticoagulants such as dabigatran / Pradaxa, the apixaban / Eliquis, rivaroxaban / Xarelto, indicated in the prevention phlebitis, embolisms or strokes in the indication atrial fibrillation;
o anti-vitamin K, very effective but little used today because require very close monitoring of coagulation.
Preferably, the additional active agent will preferably be chosen from angiotensin II receptor antagonists.
The compositions according to the present invention are likely to be administered by oral, parenteral, mucosal or cutaneous. The parenteral route includes preferably subcutaneous, intravenous, intramuscular or intraperitoneal, although the latter is rather reserved for the animal. The dirty muco pathway includes preferentially the administration by the nasal route, by the pulmonary route, by the mucosa rectally. The dermal route advantageously comprises the dermal route, in particular via a transdermal patch, typically a patch. Other ways administration especially concerning diseases affecting the nervous system Central (SNC), WO 2015/087016

25 PCT / FR2014 / 053309 especially Niemann-Pick type C disease, Parkinson's disease or disease Alzheimer's, are the intrathecal or spinal route.
The compositions according to the present invention also comprise a excipient pharmaceutically acceptable. Any suitable excipient may be used for forms galenics known to those skilled in the art, in particular with a view to administration systemic, preferentially for the oral administration of a administration parenteral administration of cutaneous or mucosal administration, particularly by subcutaneous cutaneous, intravenous, intramuscular, intraperitoneal, nasal, pulmonary, rectal, dermal, intrathecal or spinal.
There may be mentioned, for example, saline, physiological, isotonic solutions, buffered etc., compatible with pharmaceutical use and known to humans job. The compositions may contain one or more agents or selected vehicles from dispersants, solubilizers, stabilizers, preservatives, etc. Agents or vehicles can be used in formulations (liquid and / or injectable), in particular the methylcellulose, hydroxymethylcellulose, carboxymethylcellulose, polysorbate 80, mannitol, gelatin, lactose, vegetable oils, acacia, liposomes, etc. The compositions can be formulated as injectable suspensions, gels, oils, tablets, suppositories, powders, capsules, aerosols, etc.
possibly at means of galenic forms or devices providing sustained release and or delayed. For this type of formulation, an agent such as that the cellulose, carbonates or starches.
We can advantageously imagine a galenic form in which the cyclodextrin according to the invention is administered in a form complexed to a substance lipid, way to allow its passage into the bloodstream after administration by noninvasive pathways, and / or where the passage of the blood-wall barrier encephalic is desired, to allow the passage of this barrier. Of these substances lipids, one may include tocopherol.
The compositions that can be administered to an individual in the framework of the invention comprise between 1 and 100 mg / kg, preferably between 20 and 70 mg / kg, even more preferably between 30 and 50 mg / kg, and still more most preferred mg / kg of methyl-cyclodextrin as defined in the present invention, compared WO 2015/087016

26 PCT / FR2014 / 053309 to the total weight of the individual. Of course, those skilled in the art are able to adapt the dose of methyl-cyclodextrin defined in this application depending on the weight of the individual to treat.
Advantageously, the compositions are therefore capable of being administered in effective therapeutic amount in an individual suffering from an illness likely to be treated and / or prevented by an increase in HDL cholesterol, or at a individual predisposed to develop such a disease. Quantity means therapeutic effective, a sufficient amount of composition likely to be administered at a individual to prevent and / or treat a disease as defined in present invention.
The compositions used in the invention may be administered according to modalities variables. In particular, they can be injected one to five times per weeks during 1, 2, 3, 4 weeks, or even 1, 2, 3 or more months. The conditions administration include treatment at intervals of several weeks or several months.
For the treatment and / or prevention of atherosclerosis in particular, consider advantageously an administration at intervals of 3 to 6 months, which can go up to 12 month, lasting one week.
Another subject of the invention relates to the use of the compositions pharmaceutical as defined in this application to further promote a decrease in triglycerides and circulating fatty acids.
A further object of the invention is the use of the compositions pharmaceutical products as defined in this application to reduce the plates atheroma.
The use of the compositions of the invention makes it possible at the same time to increase the rate of HDL-cholesterol, lowering triglyceride and fatty acid levels circulating in the blood and reduce atheromatous plaque. The composition of the invention also allows to induce a decrease in the MMP-9 responsible for the weakening of plates of atheroma, and therefore complications related to atheroma. All these effects are obtained without changes in LDL-cholesterol levels, and without any toxicity being identified. The WO 2015/087016

27 PCT / FR2014 / 053309 The decrease in MMP-9 also suggests the interest of the composition according to the invention for use in the treatment and / or prevention of cancer.
The compositions as defined in the present application also a interest in healthy subjects. The compositions of the invention allow including reduce triglyceride levels and the concentration of free fatty acids and to increase the good cholesterol (HDL cholesterol), thereby promoting good health.
subject. It is why the present invention also relates to a non-use therapeutic methyl-cyclodextrin compositions as defined above in the present application to reduce the level of triglycerides and / or to reduce the concentration of free fatty acids and / or to increase the level of good cholesterol (HDL-Cholesterol).
Thus, the methyl-cyclodextrin composition according to the present invention may be used as nutraceutical composition, food, functional food or dietary supplement. Nutraceutical composition, food, food functional, we means a food that contains ingredients with beneficial effects for health or capable of improving physiological functions. By complement food, we means a foodstuff intended to supplement the normal diet.
Normal or healthy subjects most likely to benefit from these compositions are individuals aged 40 or over, preferably 50 years of age or over, for example having between 50 and 70 years old. These subjects do not suffer from any pathology.
The composition is intended for human food can take any shape, and in particular be a liquid, a paste or a solid. In particular, the composition maybe a dairy product such as cheese, butter, yoghurt or cream, a product based on fruit such as fruit juice, compote or fruit paste, drink, or a food solid, like a snack, a biscuit, or whatever. It can also be formulated in form tablets, a powder, a capsule, a tablet, a solution, a concentrated, syrup, suspension, or dispersion.
The following examples serve to illustrate and show other aspects and benefits of the invention and must be considered as non-limiting.

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28 PCT / FR2014 / 053309 LEGEND OF FIGURES
Figure 1: 1 H NMR spectrum of a methyl-13-cyclodextrin according to the invention.
Figure 2: Measurement of cholesterol efflux versus MS of methyl-13-cyclodextrins.
Figure 3: Expression of the ABCA1 transporter according to methylated MS
cyclodextrins.
Figure 4: Expression of the ABCG1 transporter according to methylated MS
cyclodextrins.
Figure 5: Expression of SR-B1 Receptor in MS Function of Methyl-13-cyclodextrins.
EXAMPLES
Example 1 In Vivo Study of the Methyl-Cyclodextrins of the Invention Male Apo E - / - mice, 11 weeks old, were used.
The methyl-cyclodextrin composition used is the commercial product KLEPTOSE
CRYSMEB (ROQUETTE).
The mice were distributed in 6 groups:
- 2 groups of mice subjected to a normal diet for 16 weeks. This model Atheroma is characterized by early disease. the first group was treaty with a placebo solution (control mice, n = 9), the second group was treated with 40 mg / kg KLEPTOSE CRYSMEB (treated mice, n = 12).
-2 groups of mice fed a high cholesterol diet, during 11 weeks.
This model of atheroma is characterized by an advanced disease, the first group a treated with a placebo solution (control mice, n = 9), the second group summer treated with 40 mg / kg KLEPTOSE CRYSMEB (treated mice, n = 11).
The treatments were performed by intraperitoneal injection of a volume of 200 ', IL of placebo (PBS solution) or KLEPTOSE CRYSMEB solution, three times per week.
The blood samples were taken at the sacrifice of the animals, after the 16 or 11 weeks of treatment, by cardiac puncture, to determine the impact of treatment on lipidemia.

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29 PCT / FR2014 / 053309 Other values such as blood glucose, hematocrit values (rate hemoglobin, number of erythrocytes, leucocytes, lymphocytes and platelets) were measured without no difference was observed between the different groups of mouse. By therefore, these results are not presented here.
After sacrifice, a quantification of the atheromatous surface has been realized on each group of mice by computed-assisted morphometry, out of 4 sections 101-IM aortic strains labeled with haematoxiline and Oil-ref-O, taken at 200, 400, 600 and 8001-1M after the departure of the cusp valve, according to the method described in Article Carigiuri G, Groyer E, Khaliou-Lasehet J, Ai I iaj Zen A, Sainz J, Urban D and have. Redueed iminunoregulatory T in the blood of atherosclerotie rnice with plaque thrombosis.
Artenioseler Thromb Vase Biol 2005; 25: 1659-1664. . Immunological markings histochemical MMP-9 atherosclerotic plaques were also performed and quantified to determine their fragility (R & D Systems kit).
The results obtained are shown in Tables 1 and 2.

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30 PCT / FR2014 / 053309 Table 1: Mice under normal diet and treated with a composition of methyl-cyclodextrins according to the invention Control mice (n = 9) Treated mice (n = 12) p Body weight (g) 31.84 2.70 30.71 3.09 0.4137 lipemia Total cholesterol (mmol / L) 14.49 1.65 14.19 3.02 0.8070 LDL cholesterol (mmol / L) 9.29 1.22 9.76 2.34 0.8621 HDL cholesterol (mmol / L) 2.77 0.15 3.03 0.38 0.1451 Triglycerides (mmol / L) 2.20 0.66 1.65 0.55 0.0491 Free fatty acids (mmol / L) 1.33 0.36 1.03 0.40 0.0955 Sinus of the aortic valve Oil-red-O, x 103 ium2 295.06 36.25 246.14 46.38 0.0278 MMP-9,% 6.35 4.78 3.26 4.84 0.0481 Aorta Plate area (%) 5.56 2.65 3.39 1.26 0.0278 The values are expressed as mean standard deviation.
The values of p are calculated according to the Mann-Whitney U-test.

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31 PCT / FR2014 / 053309 Table 2: Mice on a high cholesterol diet and treated with a methyl-cyclodextrin composition according to the invention Control mice (n = 9) Treated mice (n = 11) p Body weight (g) 30.44 2.34 28.34 2.80 0.1144 lipemia Total cholesterol (mmol / L) 21.48 1.37 20.17 1.37 0.4119 LDL cholesterol (mmol / L) 17.05 3.81 17.51 4.00 0.9999 HDL cholesterol (mmol / L) 3.44 0.49 4.16 0.82 0.0334 Triglycerides (mmol / L) 1.47 0.65 0.90 0.36 0.0136 Free fatty acids (mmol / L) 0.99 0.25 0.75 0.20 0.0310 Sinus of the aortic valve Oil-red-O, x 10311m2 465.56 66.99 439.45 86.58 0.4411 MMP-9,% 22.80 15.88 12.45 9.80 0.119 Aorta Plate area (%) 24.73 6.75 15.00 7.56 0.0200 The values are expressed as mean standard deviation.
The values of p are calculated according to the Mann-Whitney U-test.
It should be noted in these results that the displacement of the values resulting from the treatment is obtained not only after a rich diet, but also and so surprising and favorable as a result of a normal diet, not enriched in cholesterol.
In both cases, an increase in HDL-cholesterol was observed, showing the interest of the methyl-cyclodextrins according to the invention for a use in the treatment and / or prevention of diseases that may be treated and / or prevented by an increase in HDL cholesterol.

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32 PCT / FR2014 / 053309 In both cases, the surface of the atheromatous plaques is significantly decreased. A
significant decrease of almost 40% in the atheromatous area was observed. These results demonstrate the interest of the methyl-cyclodextrins according to the invention, for a use in the treatment and / or prevention of atherosclerosis or complications linked to an atheroma, A decrease in triglycerides, free fatty acids and MMP-9 have have been observed, and reinforce the interest of methyl-cyclodextrins according to the invention, for a use in the treatment and / or prevention of atherosclerosis or complications linked to an atheroma.
Example 2 In Vitro Study of the Methyl-Cyclodextrins of the Invention This example illustrates the effect of different cyclodextrins on efflux of cholesterol tested with aortic endothelial cells. The aortic endothelium is the tissue of the plate atheroma which is in direct contact with the blood compartment, and so in direct contact with HDL.
Equipment Cells: endothelial cells were endothelial cells aortic bovine adult (ABAE).
Cyclodextrins: the cyclodextrins used were: a 13-cyclodextrin unmethylated, hereinafter referred to as 13-CD, a methyl-13-cyclodextrin presenting an MS
0.2, hereinafter referred to as M13CD-0.2, a methyl-13-cyclodextrin presenting an MS of 0.67 (KLEPTOSE CRYSMEB, ROQUETTE FRERES), hereinafter referred to as M13CD-0.67 , a methyl-13-cyclodextrin having an MS of 1.8 (RAMEB, CYCLOLAB), hereinafter after designated M13CD-1.8.
Incubation of cells with cyclodextrins:
0.5 IJCi / mL of H3-cholesterol was incorporated into serum (10% serum horse / veal (HS / CS)) for 6 hours at 37 C in order to obtain a culture medium enriched radioactive cholesterol. After this incorporation, the culture medium has been finalized by WO 2015/087016

33 PCT / FR2014 / 053309 incorporation of DMEM (Dulbecco's Modified Eagle's medium) supplemented with 10% HS, 10% CS, bFGF (basic fibroblast growth factor) lng / mL, 2mM glutamine and Gentamicin 50p g / mL.
The cells were then cultured in this radioactive medium for 36 hours for their allow to incorporate radioactive cholesterol. The middle was then eliminated and cells rinsed twice with warm DMEM containing 0.1% serum albumin bovine (SAB).
The cells were then cultured in DMEM / bFGF 1ng / mL containing 1 mM of each cyclodextrin for 24 hours (1.4 mL / well).
A. Measurement of cholesterol captured by cyclodextrins In this example, the Applicant has studied the effect of cyclodextrins on efflux of cholesterol, depending on their degree of methylation.
Two hours after the start of incubation in the presence of each cyclodextrin, 500 ', IL
of medium were collected and the radioactive cholesterol assay cyclodextrin a was performed using a scintillation counter (Tri-carb 2100TR). The radioactivity a was measured in disintegration per minute (DPM). The capacity of each cyclodextrin to capture the cellular cholesterol was then estimated by performing the calculation next: ((1,4 mL / 0.5 mL) * DPM measured).
Values were then converted to percentages that are compared to the incubated condition without cyclodextrin (control) representing 100%.
Each experiment was performed in triplicate and repeated twice.
The results obtained are shown in Figure 2.
Results:
MI3CD-0.2 and MI3CD-0,67 according to the invention cause an efflux of cholesterol significantly lower than the I3CD and MI3CD-1,8 non-compliant with the invention.
The inventors have thus demonstrated a lower affinity with respect to the cholesterol for methylated cyclodextrins according to the invention illustrating an efflux of cholesterol Under the form of HDL-cholesterol and not in the form of cholesterol complexed with cyclodextrins, or phospholipid vesicles.

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34 PCT / FR2014 / 053309 B. Expression of carriers responsible for cholesterol efflux Intracellular: ABCA1 and ABCG1 In this example, the Applicant has studied the expression of two carriers responsible for cholesterol efflux, the ABCA1 receptor and the receptor ABCG1, following treatment with cyclodextrins of different degrees of methylation.
After 24 hours of treatment with each cyclodextrin, the cells were rinsed two times in cold PBS (8 g / L NaCl, 0.2 g / L KCl, 0.2 g / L KH2PO4, 2.87 g / L NA2HPO4 (12:20) pH 7.4) and lysed with 500 μl of lysis buffer RLT (Biorad). Three wells have been used and pooled for each condition. Purification of messenger RNAs (MRNA) has been performed using the RNeasy Total RNA Extraction kit (Qiagen) in following the manufacturer's recommendations. At the end of this purification step, the concentration of MRNA was estimated by the absorbance values obtained at 260, 280 and 320 nm. The purity of the samples was verified by carrying out the 260/280 report and only samples with a value greater than 2 were considered for our studies.
For each sample, cDNAs were synthesized from 250 ng mRNA
using the Supermix Supermix iScript Reverse Transcription Kit (Biorad) by following the instructions from the manufacturer. Real-time PCR experiments were performed in using the Sso kit Fast EvaGreen Master Mix (BioRad) and primers designed and optimized by the Applicant (Table 3).
Table 3:
MRNA Species Sequences NO
of Accession ABCA1 Bos taurus F 5'-gtgtctcgcctgttctcag-3 '(SEQ ID No. 1) NM_001024693 R 5'-gaaacatcacctcctgccg-3 '(SEQ ID No. 2) ABCG1 Homo F 5'-gaggaagaaaggatacaagacc-3 '(SEQ ID BCO29158 sapiens No 3) R 5'-gtcagtatctccttgaccattt-3 '(SEQ ID No. 4) The amplification was carried out on 40 cycles with a temperature hybridization of 60 C in using the Biorad CFX96 thermocycler. For each pair of primers, the specificity amplification was verified by performing melting curves and effectiveness has been determined by performing dilution curves. Expression levels measured have WO 2015/087016

35 PCT / FR2014 / 053309 then were normalized to the expression levels of B-actin and the cyclophilin B in using the AACt method.
Each experiment was performed in triplicate and repeated twice.
The results obtained are shown in Figures 3 and 4.
Results:
The results show that in response to treatment with cyclodextrin, way general, the expression of ABCA1 and ABCG1 receptors by the cells endothelial decreases compared to control.
In particular, the MI3CD-0,2 and MI3CD-0,67 according to the invention cause a decrease lower ABCA1 and ABCG1 receptor expression compared to I3CD and MI3CD-1.8.
By combining these results with those obtained above (point A), the inventors have showed that the I3CD and MI3CD-1,8 not in accordance with the invention induced a quick efflux and important cholesterol and a stronger reaction of the cell in decreasing sharply the expression of ABCA1 and ABCG1 receptors.
On the other hand, the MI3CD-0,2 and MI3CD-0,67 according to the invention, even if they induce lower efflux of cholesterol, decrease much less clearly expression ABCA1 and ABCG1 receptors.
The kinetics of efflux of cholesterol of methylated cyclodextrins conform to the invention is thus totally different from that of cyclodextrins which do not comply with the invention. he is postulated by the inventors that this particular kinetics of methylated cyclodextrins according to the invention has the advantage of less disturbing homeostasis of cell cholesterol.
C. Study of SR-B1 (Scavenger Receptor class B) receptor expression member 1), an HDL receptor, of endothelial cells treated with cyclodextrins In this example, the Applicant has studied the effect of cyclodextrins on expression an HDL receiver, the SR-B1 receiver, depending on their degree of methylation.
One of the roles of SR-B1 receptors in the endothelium is to promote efflux free cholesterol to the HDL from the circulation.

WO 2015/087016

36 PCT / FR2014 / 053309 The expression of the SR-B1 receptor was studied according to the protocol as described previously, using primers designed and optimized by the complainant (Table 4).
Table 4:
MRNA Species Sequences NO of Accession SCARB1 Bos taurus F 5'-cccttaatccacctcatcaatc-3 '(SEQ ID No. 5) NM_174597 R 5'-gaagtttttgacccctgtgaac-3 '(SEQ ID No. 6) Each experiment was performed in triplicate and repeated twice.
The results obtained are shown in Figure 5.
Results:
MI3CD-0,67 according to the invention causes a sharp increase in the expression of SR-B1 receptor, about 50% of basal expression.
In conclusion, the inventors have shown that methyl-cyclodextrins according to the current The invention had a low affinity for cholesterol, limited decrease in carriers responsible for the efflux of cholesterol and increased the expression of SR-B1 to HDL, which has the effect of promoting the transport of cholesterol cytoplasmic to the blood and therefore to increase cholesterol Plasma especially HDL cholesterol.
The inventors have therefore demonstrated that methyl-cyclodextrins according to the invention despite their low affinity for cholesterol did not require the addition of vesicles phospholipids to exert their effects illustrated by the results in vivo from example 1 (Tables 1 and 2).
These results reinforce the interest of methyl-cyclodextrins according to the invention, for a use in the treatment and / or prevention of to be treated and / or prevented by an increase in the HDL cholesterol level.

Claims (28)

37 1. A pharmaceutical composition comprising at least one methyl-cyclodextrin with a degree of molar substitution between 0.05 and 1.5, for its use in the treatment and / or prevention of diseases that may be treated and / or prevented by an increase in HDL cholesterol. 2. A pharmaceutical composition comprising at least one methyl-cyclodextrin with a degree of molar substitution between 0.05 and 1.5, for its use in the treatment and / or prevention of atherosclerosis or related complications atheroma, and / or diseases of the central nervous system. 3. A pharmaceutical composition for use according to claim 1 or 2, in the treatment and / or prevention of atherosclerosis or related complications atheroma. 4. Pharmaceutical composition for use according to any one of Claims 1 to 3 in the treatment and / or prevention of complications linked to a atheroma that include ischemia, preferably myocardial ischemia, diseases coronary heart disease, angina pectoris, acute coronary syndrome, infarction myocardium, mesenteric infarction, cerebrovascular accident, aneurysm or arterial disease lower limbs and their consequences. Pharmaceutical composition for its use according to claim 1 or characterized in that diseases of the central nervous system are selected among the Alzheimer's disease, Parkinson's disease, and lysosomal diseases affecting the central nervous system. Pharmaceutical composition for its use according to claim 5, characterized in that the lysosomal diseases affecting the central nervous system are chosen from Niemann-Pick disease, preferentially Niemann-Pick type disease To the Niemann-Pick type B disease, or Niemann-Pick type C disease. 7. Pharmaceutical composition for use according to any one of Claims 1 to 6, characterized in that the methyl-cyclodextrin has a degree of molar substitution between 0.2 and 1.2, preferably between 0.4 and 0.9. 8. Pharmaceutical composition for use according to any one of Claims 1 to 7, characterized in that the methyl-cyclodextrin has a degree of molar substitution between 0.6 and 0.8. 9. Pharmaceutical composition for use according to any one of Claims 1 to 8, characterized in that the methyl-cyclodextrin is a methyl-.beta.-cyclodextrin. 10. Pharmaceutical composition for use according to any one of Claims 1 to 9, characterized in that the methyl-cyclodextrin is substituted on the hydroxyl carried by the carbon C2 of the glucopyranose units, or by the C3 carbons and / or C6 of the glucopyranose units, or by a combination of the C2, C3 carbons and / or C6, preferably C2 and C6 glucopyranose units. 11. Pharmaceutical composition for use according to any one of Claims 1 to 10, characterized in that the methyl cyclodextrins comprises one or more methyl-.beta.-cyclodextrins chosen from group consisting of methyl-.beta.-cyclodextrins substituted on the hydroxyl borne by the carbon C2 of the units glucopyranoses, hydroxyl-substituted methyl-.beta.-cyclodextrins carried by C3 and / or C6 carbons of glucopyranose units, methylbeta.-substituted cyclodextrins on the hydroxyl borne by the carbons C2, C3 and / or C6, preferably C2 and C6 units glucopyranoses and said methyl- .beta.-cyclodextrins having a degree of substitution molar range from 0.6 to 0.8. 12. Pharmaceutical composition for use according to any one of Claims 1 to 11, characterized in that the methyl cyclodextrins contains at least 50, 60, or 75% methyl substituted on the hydroxyl worn by the C2 carbon of glucopyranose units. 13. Pharmaceutical composition for use according to any one of Claims 1 to 12, characterized in that the composition comprising at least least one methyl-cyclodextrin also includes a cyclodextrin, especially .beta..
cyclodextrin, unsubstituted and / or cyclodextrin, in particular .beta.
cyclodextrin substituted with sulfobutyl ether (SBE-) or hydroxypropyl (HP-) groups, preferably with a degree of molar substitution of between 0.05 and 1.5. 14. Pharmaceutical composition for use according to any one of Claims 1 to 13 further comprising at least one active agent additional. 15. Pharmaceutical composition for its use according to claim 14, characterized in that the additional active agent is an active agent used in the treatment of atherosclerosis or complications related to atheroma, preferably, - a statin, a platelet antiaggregant or anticoagulant agent, preferably selected aspirin, clopidogrel, new oral anticoagulants such as dabigatran, apixaban, rivaroxaban, and anti-vitamin K, an antihypertensive agent, preferably selected from among inhibitors angiotensin converting enzyme such as perindopril, captopril, enalapril, the lisinopril or ramipril, angiotensin II receptor antagonists as the losartan, valsartan, or candesartan, and / or among beta-blockers as acebutolol, labetalol, nadolol, oxprenolol, penbutolol, pindolol, or propranolol - or a combination of these. 16. Pharmaceutical composition for its use according to claim 15, characterized in that the additional active agent is an antagonist of receiver to angiotensin II, such as losartan, valsartan, or candesartan. 17. A pharmaceutical composition for use according to any one of Claims 1 to 16, characterized in that the composition is to be administered by oral, parenteral, or dermal administration or mucosal. 18. Pharmaceutical composition for use according to any one of claims 1 to 17 further promoting a decrease in the rate of triglycerides and of circulating fatty acids. 19. Pharmaceutical composition for use according to any one of Claims 1 to 4 and 7 to 17 for reducing and / or preventing plates atheroma. 20. Composition for use according to any one of the claims 1 to 19, characterized in that said composition is free of vesicles of phospholipids. 21. Non-therapeutic use of a composition comprising at least one methyl cyclodextrin having a degree of molar substitution of between 0.05 and 1.50, preferably between 0.60 and 0.80, to decrease the level of triglycerides and / or decrease the concentration of free fatty acids and / or to increase the rate of cholesterol-HDL. 22. Use according to claim 21, characterized in that the methyl-cyclodextrin has a degree of molar substitution of between 0.2 and 1.2, preferentially between 0.4 and 0.9. 23. Use according to claim 21 or 22, characterized in that the methyl cyclodextrin has a degree of molar substitution of between 0.6 and 0.8. 24. Use according to any one of claims 21 to 23, characterized in that the methyl-cyclodextrin is a methyl-beta-cyclodextrin. 25. Use according to any one of claims 21 to 24, characterized in that the methyl-cyclodextrin is substituted on the hydroxyl carried by carbon C2 units glucopyranoses, or by the C3 and / or C6 carbons of the glucopyranose units, or by one combination of the C2, C3 and / or C6 carbons, preferably C2 and C6 of the units glucopyranose. 26. Use according to any one of claims 21 to 25, characterized in that the methyl-cyclodextrin composition comprises one or more methyl-.beta.-cyclodextrins selected from the group consisting of methylbeta.-cyclodextrins substituted on the hydroxyl carried by the C2 carbon of the glucopyranose units, methyl .beta.-cyclodextrins substituted on the hydroxyl borne by C3 carbons and / or C6 units glucopyranoses, hydroxyl-substituted methyl-.beta.-cyclodextrins carried by C2, C3 and / or C6, preferably C2 and C6, of the glucopyranose units and said methyl- .beta.-cyclodextrins having a molar degree of substitution between 0.6 and 0.8. 27. Use according to any one of claims 21 to 26, characterized in that the Methyl-cyclodextrin composition comprises at least 50, 60, or 75% of methylated substituted on the hydroxyl carried by the C2 carbon of the glucopyranose units. 28. Use according to any one of claims 21 to 27, characterized in that the composition comprising at least one methyl-cyclodextrin also comprises a cyclodextrin, in particular .beta. ¨cyclodextrin, unsubstituted and / or cyclodextrin, in particular .beta. ¨cyclodextrin substituted by sulfobutyl ether groups (SBE-) or hydroxypropyl (HP-), preferably with a molar degree of substitution between 0.05 and 1.5.